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Principal Investigator: Margaret M. Billingsley
Organization: UNIVERSITY OF PENNSYLVANIA
Fiscal Year: 2021
Award: $46,036
Funding agency: National Cancer Institute
PROJECT SUMMARY
CAR T cell immunotherapy is FDA approved for the treatment of acute lymphoblastic leukemia (ALL) and large
B cell lymphoma and has shown success in inducing durable remission. However, the therapy is also associated
with causing severe, life-threatening side effects—including cytokine release syndrome, B cell aplasia, and
neurotoxicity—in 70% of patients receiving the treatment. Thus, there is a need to develop CAR T cells that
maintain their therapeutic efficacy while minimizing adverse effects. Currently, CAR T cells are engineered using
viruses that induce permanent CAR expression, but investigations into mRNA-based CAR T cells—which result
in transient CAR expression—have been utilized in clinical trials and shown potential for mitigating long-term
side effects of the immunotherapy. To create these mRNA CAR T cells, electroporation is utilized for T cell
transfection, but it is cytotoxic and has no potential for translation to in vivo T cell delivery. Thus, this investigation
aims to explore ionizable lipid nanoparticles (LNPs) as a delivery tool for the ex vivo engineering of T cells. LNPs
have shown potent mRNA delivery in various cell types and can be easily modified to alter the physicochemical
properties that impact delivery, which will allow for their optimization as a delivery platform for T cells specifically.
In Aim 1, 24 novel LNPs will be screened for their ability to functionally deliver mRNA with low toxicity, and the
top-performing LNP will be further optimized to determine the best formulation for delivery to primary T cells. In
Aim 2, the LNP formulation selected in Aim 1 will be used to encapsulate CAR mRNA with different modifications
to determine the best mRNA cargo for LNP-based delivery to T cells. With the top LNP and CAR mRNA cargo
selected, Aim 3 will validate LNPs as a method for CAR T cell engineering as compared to electroporated mRNA-
CAR T cells and virus-based CAR T cells via a survival study using an ALL mouse model. The completion of
these aims will identify and optimize a mRNA delivery platform for T cells that, in future investigations, can be
utilized for the screening of new CAR constructs or in vivo delivery. Ultimately, this work—conducted as an
interdisciplinary project between sponsors in the Bioengineering Department and Medical School at University
of Pennsylvania—will allow for the development of a novel LNP delivery platform for immune cell engineering.
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